CN105671157A - Kit for synchronous detection of animal origin ingredients in meat and meat product and application of kit - Google Patents
Kit for synchronous detection of animal origin ingredients in meat and meat product and application of kit Download PDFInfo
- Publication number
- CN105671157A CN105671157A CN201610107349.4A CN201610107349A CN105671157A CN 105671157 A CN105671157 A CN 105671157A CN 201610107349 A CN201610107349 A CN 201610107349A CN 105671157 A CN105671157 A CN 105671157A
- Authority
- CN
- China
- Prior art keywords
- meat
- kit
- enzyme
- sample
- detection
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/686—Polymerase chain reaction [PCR]
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Analytical Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biophysics (AREA)
- Immunology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Physics & Mathematics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a kit for synchronous detection of animal origin ingredients in meat and a meat product and application of the kit, belonging to the technical field of molecular biology identification/detection of the meat and the meat products. The detection principle of the kit comprises: extracting genomic DNA (deoxyribonucleic acid) in a sample to be detected, amplifying a target fragment (710bp) in mitochondrial DNA COI segment of the sample by using a single universal primer, adopting 1 percent of agarose gel electrophoresis to detect an amplification result after amplifying, if the result is consistent with the target fragment, performing restriction enzyme digestion on the amplified target fragment by using specific restriction endonuclease to cut the amplified target fragment into species specificity DNA fragments with different sizes, adopting 3 percent of agarose gel electrophoresis to analyze the restriction enzyme digestion result, and judging the species composition in the sample according to characteristic restriction enzyme digestion strips. The kit has the advantages that the operation is simple, the accuracy and reliability can be realized, the detection time is short, the cost performance is high, multi-species can be simultaneously identified, and the like, and the kit can be widely popularized and applied in the field of animal food safety inspection.
Description
Technical field
The present invention relates to the test kit of animal derived materials in a kind of synchronous detection SDS in broiler chickens, also relate to the application of this test kit simultaneously, belong to SDS in broiler chickens molecular biology discriminating/detection technique field.
Background technology
Bread is the staff of life, and food, with An Weixian, eats the important aspect that relieved healthy green meat product is food quality safety. But in animal derived raw material and converted products, the phenomenon that animal component is adulterated, fake, adulterate and be not intended to pollution often occurs. A lot of meat adulteration events that domestic market exposes in the recent period have caused the public to the worry of food safety, even if in the Europe having the strictest food safety system in the world, and the fakement phenomena also occurring hanging out a sheep's head but actually sell dog's meat unavoidably.
Beef is the meat-based food that the people of the world dotes on, and is rich in amino acid, vitamins B12, the nutritive ingredient such as ferro element, and protein content height in beef, lipid content is low, and taste is very delicious, enjoys the laudatory title of " in meat tender son ". At present, on market, the price of beef is generally higher than other meat, and this also makes some illegal retailers adulterate under the ordering about of profit, replaces high quality beef with beef cheap, inferior and sells, even some retailers directly hang out a sheep's head but actually sell dog's meat, and pretend to be beef to sell with other meats.
The adulterated of meat relates to all many-sides such as food safety, public health, religion, people's life is had great negative impact, not only seriously violates the criterion of meat market fair competition, also can seriously affect the physical and mental health of human consumer, infringement consumer rights. Emerge in an endless stream instantly in meat adulteration form, develop and apply and fast, accurately can differentiate that the new technology of animal derived materials has become the research focus of field of food safety.
Traditional animal derived materials detection method relates to the numerous areas technology such as anatomy, histology, chemistry, biological chemistry, spectrum, chromatogram, immunology, comprise the Protocols in Molecular Biology based on nucleic acid (such as PCR, real-time quantitative PCR, molecular fingerprint technology etc.), immuno analytical method based on protein molecular structure, infrared spectrum technology and mass-spectrometric technique etc., but all show certain limitation.The multiple PCR primer system of a kind of pig sheep ox animal derived materials as disclosed in the patent of invention of publication No. CN105177150A, comprise the primer pair I for pig 12SrDNA gene design, for the primer pair II of sub-base III gene design of sheep cytochrome c oxidase, for the primer pair III of ox TNFRSF10A gene design, detection method comprises: taking sample total DNA to be checked as template, utilize primer I, II, III carries out multiplex PCR amplification, increase complete gel electrophoresis result of determination, amplify 290bp band and namely comprise pig derived component, 370bp band comprises sheep derived material, 190bp band comprises calf-derived Cyclospora, the method is easy and simple to handle, highly sensitive, high specificity, pig sheep ox 3 kinds of animal derived materials in sample can be made detection simultaneously analyze, but primer proportioning there is is certain requirement, and identify the limited amount of species, operate loaded down with trivial details and detected result poor accuracy.
In recent years, based on the development of DNA molecular technology, the species identification method superiority based on DNA especially Mitochondrial DNA highlights. Mitochondrial DNA is at high temperature stablized, and it is prevalent in most cells and tissue, by individual morphology feature limits, (sampling amount is not little, sample damage does not affect recognition result), also do not affect by soil individual development, it is convenient to extract and obtains (being as good as with conventional genome DNA extracting method), be widely used in biological study. DNA bar code technology based on Mitochondrial cytochrome c oxidase subunit gene I (CO I) is widely used in zooscopy, the standard fragment adopted is the section of DNA in mitochondrial COⅠ gene, carries out identifying fast and accurately and qualification to species by this standard fragment. But, the not yet application in SDS in broiler chickens animal derived materials detects of this technology.
Summary of the invention
It is an object of the invention to provide the test kit of animal derived materials in a kind of synchronous detection SDS in broiler chickens.
Meanwhile, the present invention reoffers the application in animal derived materials in synchronous detection SDS in broiler chickens of a kind of mentioned reagent box.
In order to realize above object, the technical solution adopted in the present invention is:
The test kit of animal derived materials in synchronous detection SDS in broiler chickens, comprises Hpa II enzyme and primer pair H/L;
Primer pair H/L is:
LCO1490:5 '-GGTCAACAAATCATAAAGATATTGG-3 ',
HCO2198:5 '-TAAACTTCAGGGTGACCAAAAAATCA-3 '.
Concrete, this test kit can also comprise TaqMasterMix (containing dyestuff), NEBuffer, DNAMarker, deionized water etc.
The application in animal derived materials in synchronous detection SDS in broiler chickens of mentioned reagent box, comprise: taking comprise CO I gene testing sample DNA as template, primer pair H/L is utilized to carry out pcr amplification, amplified production is gel electrophoresis analysis after Hpa II enzyme digestions, according to characteristic enzyme slitting band result of determination;
Described primer pair H/L is:
LCO1490:5 '-GGTCAACAAATCATAAAGATATTGG-3 ',
HCO2198:5 '-TAAACTTCAGGGTGACCAAAAAATCA-3 '.
The reaction system of described pcr amplification is: 2 × TaqMasterMix (containing dyestuff) 5 μ L, 10ng/ μ L template DNA 1 μ L, each 0.2 μ L of 10pmol/L primer LCO1490, HCO2198, deionized water 3.6 μ L, cumulative volume 10 μ L.
The response procedures of described pcr amplification is: 94 DEG C of denaturation 5min; 94 DEG C of sex change 30s, 40 DEG C of annealing 30s, 72 DEG C extend 60s, 45 circulations;72 DEG C extend 10min.
The reaction system of described digestions is: above-mentioned pcr amplification product 3 μ L, 10units/ μ LHpa II enzyme 0.5 μ L, 10 × NEBuffer1.5 μ L, deionized water 5 μ L, cumulative volume 10 μ L.
The reaction conditions of described digestions is: at 37 DEG C, enzyme cuts 15min.
The corresponding relation of described characteristic enzyme slitting band and animal derived materials is: ox, 500bp, 70bp; Sheep, 500bp, 400bp, 230bp, 120bp, 80bp; Pig, 350bp, 240bp, 120bp; Chicken, 350bp, 240bp; Crow chicken, 450bp, 350bp, 240bp; Fish, 280bp, 200bp, 120bp; Mouse, 380bp, 350bp; Duck, 350bp, 280bp, 230bp.
The useful effect of the present invention:
In the present invention, in test kit synchronous detection SDS in broiler chickens, the principle of animal derived materials is: extract detected sample genomic dna, by the object fragment (710bp) in single general-purpose primer amplification sample wire mitochondrial DNA CO I section, 1% agarose gel electrophoresis detection amplification is adopted after amplification, result display is consistent with object fragment band, with specific restriction enzyme, its enzyme is cut to the species specificity DNA fragmentation of different size again, adopt 3% agarose gel electrophoresis to analyze enzyme and cut result, judge species composition in sample according to characteristic enzyme slitting band. If beef sample occurs the characteristic enzyme slitting band of other species, namely doped with other meat in expression beef, the object that detection beef is adulterated can be reached.
The present invention utilizes DNA bar code technology (DNABarcoding), for the characteristic of chondriogen high conservative, use single general-purpose primer amplification and single digestion with restriction enzyme, by amplification and enzyme being cut electrophoresis detection and the analysis of result, can be accurate, fast animal derived materials in SDS in broiler chickens is detected, have easy and simple to handle, accurately and reliably, detection time short (about 8h obtains detected result), cost performance height, the advantages such as several species discriminating simultaneously, can in animal-derived food product (comprising cooked meat product) safety detection field wide popularization and application.
Accompanying drawing explanation
Fig. 1 is the agarose gel electrophoresis figure of PCR primer detection in test example;
Fig. 2 is the agarose gel electrophoresis figure that single meat sample PCR primer enzyme cuts result detection;
Fig. 3 is the agarose gel electrophoresis figure that mixing meat sample PCR primer enzyme cuts result detection;
Fig. 4 is the mixing meat sample PCR primer agarose gel electrophoresis figure for sensitivity technique;
Fig. 5 is the mixing meat sample PCR primer digestion products agarose gel electrophoresis figure for sensitivity technique;
Fig. 6 is the agarose gel electrophoresis figure of commercially available cold fresh meat PCR primer detection;
Fig. 7 is the agarose gel electrophoresis figure that commercially available cold fresh meat PCR primer enzyme cuts result detection.
Embodiment
The present invention is only described in further detail by following embodiment, but does not form any limitation of the invention.
Embodiment 1
The test kit of animal derived materials in synchronous detection SDS in broiler chickens, comprises Hpa II enzyme and primer pair H/L;
Primer pair H/L is:
LCO1490:5 '-GGTCAACAAATCATAAAGATATTGG-3 ',
HCO2198:5 '-TAAACTTCAGGGTGACCAAAAAATCA-3 '.
Embodiment 2
The test kit of animal derived materials in synchronous detection SDS in broiler chickens, comprises Hpa II enzyme 2000units, 10pmol/L each 5mL of primer LCO1490, HCO2198,2 × TaqMasterMix (containing dyestuff) 50mL, 10 × NEBuffer30mL, DNAMarker and deionized water 100mL;
Primer pair H/L is:
LCO1490:5 '-GGTCAACAAATCATAAAGATATTGG-3 ',
HCO2198:5 '-TAAACTTCAGGGTGACCAAAAAATCA-3 '.
Test example
One, sensing range analysis
1, the extraction of material preparation and genomic dna, detection
1) extraction of genomic dna
Taking the mixing meat sample of the single meat sample of beef, mutton, pork, chicken, blackbone Chicken, the flesh of fish, rat meat, duck and ox/sheep, ox/pig, ox/chicken, ox/fish, ox/sheep/pig, sheep/pig etc. as material, it is saved to DNA extraction at putting 20 DEG C, adopt universal pillar genome to extract test kit (be century bio tech ltd purchased from health) and extract genomic dna, concrete operation carries out according to test kit specification sheets, and the genomic dna of extraction saves backup at 4 DEG C;
2) trace spectrophotometer detection genomic dna concentration
Trace spectrophotometer (ThermoScientificNanoDrop2000) is adopted to measure DNA sample concentration and preserve mensuration record, if OD260/OD280Being greater than 1.8, in interpret sample, DNA purity is higher, extracts quality good;
3) electrophoresis detection DNA quality
Adopting 1% agarose gel electrophoresis detection DNA quality, detect complete taking-up 10 μ LDNA sample, for subsequent use at being placed in 4 DEG C, all the other preserve at depositing in-20 DEG C;
2, sample detection
1) single general-purpose amplimer synthesis
Taking the general primer of invertebrates plastosome of document record as with reference to (FolmerO, BlackM, HoehW, etal.DNAprimersforamplificationofmitochondrialcytochrome coxidasesubunitIfromdiversemetazoaninvertebrates. [J] .MolecularMarineBiology&Biotechnology, 1994,3 (5): 294--299.), entrusting raw work biotechnology (Shanghai) limited-liability company synthetic primer to H/L, primer sequence is as shown in SEQIDNO:1,2;
2) pcr amplification
Taking the genomic dna of said extracted as template, utilizing primer pair H/L to carry out pcr amplification, amplification reaction system sees the following form 1;
Table 1PCR amplification reaction system
The response procedures of pcr amplification is: 94 DEG C of denaturation 5min; 94 DEG C of sex change 30s, 40 DEG C of annealing 30s, 72 DEG C extend 60s, 45 circulations; 72 DEG C extend 10min;
3) PCR primer detection
Amplified reaction is complete, get amplified production and carry out 1% agarose gel electrophoresis detection, electrophoretic band size is consistent (see Fig. 1 with the 710bp of expection, in figure, M is 2000bpMarker, N is beef sample, Y is meat samples, Z is pork sample, J is chicken meat sample, WJ is blackbone Chicken sample, F is flesh of fish sample, R is rat meat sample, D is duck sample, NY is each 50% biased sample of ox/mutton, NZ is each 50% biased sample of ox/pork, NYZ is each 1/3 biased sample of ox/sheep/pork, NJ is each 50% biased sample of ox/chicken), follow-up enzyme can be carried out and cut detection,
4) restriction enzyme Hpa II enzyme is cut
Utilizing single restriction enzyme Hpa II enzyme to cut pcr amplification product, endonuclease reaction system sees the following form 2;
Table 2 endonuclease reaction system
Endonuclease reaction condition is: at 37 DEG C, enzyme cuts 15min;
5) enzyme cuts result detection
Endonuclease reaction is complete, gets digestion products and carries out 3% agarose gel electrophoresis detection;
6) enzyme cuts result and analysis
Single meat sample enzyme is cut and be the results are shown in Figure 2 (in figure, M is 500bpMarker, NG is negative control, N is beef sample, Y is meat samples, Z is pork sample, J is chicken meat sample, WJ is blackbone Chicken sample, F is flesh of fish sample, R is rat meat sample, D is duck sample), mixing meat sample enzyme is cut and be the results are shown in Figure 3 (in figure, M is 500bpMarker, N is beef sample, Y is meat samples, Z is pork sample, J is chicken meat sample, F is flesh of fish sample, NY is each 50% biased sample of ox/mutton, NZ is each 50% biased sample of ox/pork, NJ is each 50% biased sample of ox/chicken, NF is ox/oppress each 50% biased sample, NYZ is each 1/3 biased sample of ox/sheep/pork, YZ is each 50% biased sample of sheep/pork, D is duck sample), characteristic enzyme slitting band and animal derived materials corresponding relation see the following form 3.
The corresponding relation of table 3 characteristic enzyme slitting band and animal derived materials
Two, detection method sensitivity analysis
With known beef and pork, according to different ratios mixing, (each biased sample total amount is 30mg, W/W), genomic dna is extracted with reference to method in test example one, carry out pcr amplification, with 1% agarose gel electrophoresis detection amplification (see Fig. 4 after having increased, in figure, M is 2000bpMarker, each ratio is respectively the blending ratio of pig/beef, NG is negative control), beef and pork mixing meat sample pcr amplification product is cut again with single restriction enzyme Hpa II enzyme, 3% agarose gel electrophoresis technology detection enzyme is utilized to cut result (see Fig. 5, in figure, M is 2000bpMarker, each ratio is respectively the blending ratio of pig/beef).
Embodiment 3
Test kit application in animal derived materials in synchronous detection SDS in broiler chickens in embodiment 2, comprising:
1) adopt universal pillar genome to extract genomic dna that test kit (be century bio tech ltd purchased from health) extracts commercially available cold fresh meat sample (inspect by random samples at random from certain general merchandise store and obtain), 4 DEG C save backup;
2) taking above-mentioned genomic dna as template, utilizing primer pair H/L to carry out pcr amplification (primer sequence is as shown in SEQIDNO:1,2), amplification reaction system and program are as follows:
Reaction system is: 2 × TaqMasterMix (containing dyestuff) 5 μ L, 10ng/ μ L template DNA 1 μ L, each 0.2 μ L of 10pmol/L primer LCO1490, HCO2198, deionized water 3.6 μ L, cumulative volume 10 μ L;
Response procedures is: 94 DEG C of denaturation 5min; 94 DEG C of sex change 30s, 40 DEG C of annealing 30s, 72 DEG C extend 60s, 45 circulations; 72 DEG C extend 10min;
3) amplified reaction is complete, get amplified production and carry out 1% agarose gel electrophoresis detection, electrophoretic band size is consistent (see Fig. 6 with the 710bp of expection, in figure, M is 2000bpMarker, 1～16 is commercially available cold fresh meat sample, NG is negative control), then carry out the process of Hpa II enzyme digestions, endonuclease reaction system and condition are as follows:
The enzyme system of cutting is: above-mentioned pcr amplification product 3 μ L, 10units/ μ LHpa II enzyme 0.5 μ L, 10 × NEBuffer1.5 μ L, deionized water 5 μ L, cumulative volume 10 μ L;
Enzyme slitting part is: at 37 DEG C, enzyme cuts 15min;
4) endonuclease reaction is complete, get digestion products and carry out 3% agarose gel electrophoresis detection (see Fig. 7, in figure, M is 500bpMarker, 1～12 is commercially available cold fresh meat sample, NG is negative control), as can be seen from the figure, enzyme is cut result and is comprised 500bp, 350bp, 240bp, 120bp, 70bp characteristic enzyme slitting band, judges to comprise beef and pork content in commercially available cold fresh meat sample.
Claims (9)
1. the test kit of animal derived materials in synchronous detection SDS in broiler chickens, it is characterised in that: comprise Hpa II enzyme and primer pair H/L;
Primer pair H/L is:
LCO1490:5 '-GGTCAACAAATCATAAAGATATTGG-3 ',
HCO2198:5 '-TAAACTTCAGGGTGACCAAAAAATCA-3 '.
2. test kit according to claim 1, it is characterised in that: test kit also comprises TaqMasterMix, NEBuffer, DNAMarker and deionized water.
3. test kit application in animal derived materials in synchronous detection SDS in broiler chickens as claimed in claim 1 or 2.
4. application according to claim 3, it is characterized in that: comprise the following steps: taking comprise CO I gene testing sample DNA as template, utilizing primer pair H/L to carry out pcr amplification, amplified production is gel electrophoresis analysis after Hpa II enzyme digestions, according to characteristic enzyme slitting band result of determination;
Described primer pair H/L is:
LCO1490:5 '-GGTCAACAAATCATAAAGATATTGG-3 ',
HCO2198:5 '-TAAACTTCAGGGTGACCAAAAAATCA-3 '.
5. application according to claim 4, it is characterized in that: the reaction system of described pcr amplification is: 2 × TaqMasterMix5 μ L, 10ng/ μ L template DNA 1 μ L, each 0.2 μ L of 10pmol/L primer LCO1490, HCO2198, deionized water 3.6 μ L, cumulative volume 10 μ L.
6. application according to claim 5, it is characterised in that: the response procedures of described pcr amplification is: 94 DEG C of denaturation 5min; 94 DEG C of sex change 30s, 40 DEG C of annealing 30s, 72 DEG C extend 60s, 45 circulations; 72 DEG C extend 10min.
7. application according to claim 6, it is characterised in that: the reaction system of described digestions is: amplified production 3 μ L, 10units/ μ LHpa II enzyme 0.5 μ L, 10 × NEBuffer1.5 μ L, deionized water 5 μ L, cumulative volume 10 μ L.
8. application according to claim 7, it is characterised in that: the reaction conditions of described digestions is: at 37 DEG C, enzyme cuts 15min.
9. application according to claim 4, it is characterised in that: the corresponding relation of described characteristic enzyme slitting band and animal derived materials is: ox, 500bp, 70bp; Sheep, 500bp, 400bp, 230bp, 120bp, 80bp; Pig, 350bp, 240bp, 120bp; Chicken, 350bp, 240bp; Crow chicken, 450bp, 350bp, 240bp; Fish, 280bp, 200bp, 120bp; Mouse, 380bp, 350bp; Duck, 350bp, 280bp, 230bp.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610107349.4A CN105671157A (en) | 2016-02-25 | 2016-02-25 | Kit for synchronous detection of animal origin ingredients in meat and meat product and application of kit |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610107349.4A CN105671157A (en) | 2016-02-25 | 2016-02-25 | Kit for synchronous detection of animal origin ingredients in meat and meat product and application of kit |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN105671157A true CN105671157A (en) | 2016-06-15 |
Family
ID=56306148
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610107349.4A Pending CN105671157A (en) | 2016-02-25 | 2016-02-25 | Kit for synchronous detection of animal origin ingredients in meat and meat product and application of kit |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105671157A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106048034A (en) * | 2016-07-01 | 2016-10-26 | 北京市食品安全监控和风险评估中心 | Method for scanning and analyzing animal-source components in food based on PCR-RLFP-DHPLC technology |
| CN106967802A (en) * | 2017-03-28 | 2017-07-21 | 上海上药第生化药业有限公司 | Differentiate the kit and its method and primer pair of animal derived materials in protide bio feedstocks crude product |
| CN108456736A (en) * | 2018-05-11 | 2018-08-28 | 北华大学 | Medulla Bovis seu Bubali DNA identification kits and identification method |
| CN109337992A (en) * | 2018-11-23 | 2019-02-15 | 山东省农业科学院农产品研究所 | The method of duck is adulterated in a kind of identification mutton |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105177150A (en) * | 2015-09-29 | 2015-12-23 | 上海市农业科学院 | Multiple-PCR primer system for quickly testing animal origin ingredients of pigs, sheep and cows and testing method |
-
2016
- 2016-02-25 CN CN201610107349.4A patent/CN105671157A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105177150A (en) * | 2015-09-29 | 2015-12-23 | 上海市农业科学院 | Multiple-PCR primer system for quickly testing animal origin ingredients of pigs, sheep and cows and testing method |
Non-Patent Citations (2)
| Title |
|---|
| NADIA HAIDER等: "Identification of meat species by PCR-RFLP of the mitochondrial COI gene", 《MEAT SCIENCE》 * |
| 邱德义等: "DNA条形码技术在肉品防欺诈鉴别中的应用", 《MEAT RESEARCH》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106048034A (en) * | 2016-07-01 | 2016-10-26 | 北京市食品安全监控和风险评估中心 | Method for scanning and analyzing animal-source components in food based on PCR-RLFP-DHPLC technology |
| CN106967802A (en) * | 2017-03-28 | 2017-07-21 | 上海上药第生化药业有限公司 | Differentiate the kit and its method and primer pair of animal derived materials in protide bio feedstocks crude product |
| CN108456736A (en) * | 2018-05-11 | 2018-08-28 | 北华大学 | Medulla Bovis seu Bubali DNA identification kits and identification method |
| CN109337992A (en) * | 2018-11-23 | 2019-02-15 | 山东省农业科学院农产品研究所 | The method of duck is adulterated in a kind of identification mutton |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Böhme et al. | Recent applications of omics-based technologies to main topics in food authentication | |
| Ali et al. | Multiplex PCR in species authentication: probability and prospects—a review | |
| Ballin | Authentication of meat and meat products | |
| Ha et al. | Identification of pork adulteration in processed meat products using the developed mitochondrial DNA-based primers | |
| CN104946788A (en) | PCR primer and kit for identifying eight kinds of animal-derived components | |
| CN105177150B (en) | A kind of the multiple PCR primer system and detection method of quick detection pig sheep ox animal derived materials | |
| Amaral et al. | Advances in authenticity testing for meat speciation | |
| CN105671157A (en) | Kit for synchronous detection of animal origin ingredients in meat and meat product and application of kit | |
| CN105274246B (en) | The detection kit of several species kind derived components in calf-derived Cyclospora identification and its product | |
| CN104946790B (en) | A kind of PCR method for 8 kinds of animal derived materials of identification of tracing to the source | |
| CN111334585B (en) | Primer and kit for simultaneously detecting 8 animal components, detection method and application | |
| Zhao et al. | Direct lysis-multiplex polymerase chain reaction assay for beef fraud substitution with chicken, pork and duck | |
| Rea et al. | Species identification in anchovy pastes from the market by PCR-RFLP technique | |
| Gargouri et al. | PCR–RFLP and species-specific PCR efficiency for the identification of adulteries in meat and meat products | |
| CN105506167A (en) | Method for synchronously detecting animal-origin ingredients in meat and meat products | |
| Zhao et al. | Development of a species-specific PCR coupled with lateral flow immunoassay for the identification of goose ingredient in foods | |
| CN106521016B (en) | The real-time fluorescence PCR detection method of calf-derived Cyclospora in food and feed is detected using single-copy nuclear gene | |
| CN103525908A (en) | Method for rapidly detecting chicken, duck and pig blood components in blood jelly | |
| Čapla et al. | Procedures for the identification and detection of adulteration of fish and meat products. | |
| CN104962650B (en) | A kind of synchronous PCR method and kit for differentiating animal derived materials | |
| CN103451304A (en) | Primer probe and method for performing real-time fluorescent polymerase chain reaction (PCR) detection on origin ingredients of raccoon | |
| Chappalwar et al. | Recent novel techniques applied for detection of meat adulteration and fraudulent practices | |
| CN104789692A (en) | Primer pair and kit for identifying cattle and sheep porcine-derived component | |
| CN106995852B (en) | Real-time fluorescence PCR detection method for detecting sheep-derived components in food and feed by using single-copy nuclear gene | |
| CN107012247B (en) | Real-time fluorescence PCR detection method for detecting goat-derived ingredients in food and feed by using single-copy nuclear gene |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20160615 |